Rapid Diagnosis of Drug Resistance to Fluoroquinolones, Amikacin, Capreomycin, Kanamycin and Ethambutol Using Genotype MTBDRsl Assay: A Meta-Analysis

Yuan, Feng; Liu, Sijun; Wang, Qungang; Wang, Liang; Tang, Shaowen; Wang, Jianming; Lü, Wei

doi:10.1371/journal.pone.0055292

Cited by 64 publications

(50 citation statements)

References 51 publications

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“…The line probe assay, MTBDRplus version 2 (v2) (Hain Lifescience, Nehren, Germany), detects INH and RIF resistance, and the MTBDRsl (Hain Lifescience, Nehren, Germany) is designed to detect FQ, SLID, and ethambutol resistance. However, the reported sensitivities of these assays remain less than ideal for INH and SLID (14)(15)(16).…”

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confidence: 99%

MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

et al. 2016

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e Accurate identification of drug-resistant Mycobacterium tuberculosis is imperative for effective treatment and subsequent reduction in disease transmission. Line probe assays rapidly detect mutations associated with resistance and wild-type sequences associated with susceptibility. Examination of molecular-level performance is necessary for improved assay result interpretation and for continued diagnostic development. Using data collected from a large, multisite diagnostic study, probe hybridization results from line probe assays, MTBDRplus and MTBDRsl, were compared to those of sequencing, and the diagnostic performance of each individual mutation and wild-type probe was assessed. Line probe assay results classified as resistant due to the absence of wild-type probe hybridization were compared to those of sequencing to determine if novel mutations were inhibiting wild-type probe hybridization. The contribution of absent wild-type probe hybridization to the detection of drug resistance was assessed via comparison to a phenotypic reference standard. In our study, mutation probes demonstrated significantly higher specificities than wild-type probes and wild-type probes demonstrated marginally higher sensitivities than mutation probes, an ideal combination for detecting the presence of resistance conferring mutations while yielding the fewest number of false-positive results. The absence of wild-type probe hybridization without mutation probe hybridization was determined to be primarily the result of failure of mutation probe hybridization and not the result of novel or rare mutations. Compared to phenotypic culture-based drug susceptibility testing, the absence of wild-type probe hybridization without mutation probe hybridization significantly contributed to the detection of phenotypic rifampin and fluoroquinolone resistance with negligible increases in false-positive results. While the number of incident cases of tuberculosis (TB) has been steadily declining over the past decade, the prevalence of drug-resistant disease threatens to reverse these declines (1, 2). The World Health Organization (WHO) estimated that, in 2014, 3.3% of new and 20% of previously treated TB cases were multidrug-resistant (MDR) or were resistant to isoniazid (INH) and rifampin (RIF) (3). Furthermore, 9.7% of these MDR cases were extensively drug resistant (XDR) or were resistant to at least one fluoroquinolone (FQ) in addition to a second-line injectable drug (SLID), amikacin (AMK), kanamycin (KAN), or capreomycin (CAP). Although detection of M/XDR-TB is imperative for effective disease management, less than half of all MDR-TB cases were detected in 2014 (2-4). This detection gap is due, in part, to the lack of low-cost, rapid, and accurate diagnostic technologies for M/XDR-TB (4-6).Conventional culture-based phenotypic drug susceptibility testing (DST) methods can take months to yield results (7). Molecular-based tests, however, can produce results in Ͻ1 day (5,(8)(9)(10)(11)(12). The WHO has endorsed line probe hybridization and mole...

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MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

et al. 2016

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“…Eventually, several assessments have resulted in very different results. Thus, sensitivity was 57.1 to 100% for the detection resistance in fluoroquinolone and 42 to 100% for second-line injectable drugs (SILDs) [5]. The specificity varied meanwhile from 77.3 to 100 % for fluoroquinolones and from 47 to 100 % for SLIDs [5].…”

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confidence: 99%

“…In the other hand, the test was characterized, in addition to a very average specificity, by a low sensitivity for the detection of resistance to this drug [5].…”

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confidence: 99%

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Molecular Characterization of the Resistance of <i>Mycobacterium tuberculosis</i> to Second Line Drugs in Côte d’Ivoire

Ouassa¹,

Loukou²,

Dotia³

et al. 2014

Trop. J. Pharm Res

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“…Even when available, culture-based susceptibility testing can take from weeks to months to complete. M. tuberculosis may also be tested for drug resistance using the alternative rapid, sensitive, and safer genotypic assays, which detect resistance by identifying mutations known to confer resistance to the first-and second-line drugs in a majority of clinical strains (14)(15)(16)(17)(18)(19)(20)(21)(22)(23) and minimizes the biohazard. Genotypic testing approaches that can be reduced to a few manual steps are more amenable to use at the point of care, which can dramatically expand their availability to medically underserved populations.…”

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confidence: 99%

Detection of Isoniazid-, Fluoroquinolone-, Amikacin-, and Kanamycin-Resistant Tuberculosis in an Automated, Multiplexed 10-Color Assay Suitable for Point-of-Care Use

Chakravorty

Roh

Glass³

et al. 2017

J Clin Microbiol

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Extensively drug-resistant (XDR) tuberculosis (TB) cannot be easily or quickly diagnosed. We developed a rapid, automated assay for the detection of XDR-TB plus resistance to the drug isoniazid (INH) for point-of-care use. Using a simple filter-based cartridge with an integrated sample processing function, the assay identified a wide selection of wild-type and mutant sequences associated with XDR-TB directly from sputum. Four new large-Stokes-shift fluorophores were developed. When these four Stokes-shift fluorophores were combined with six conventional fluorophores, 10-color probe detection in a single PCR tube was enabled. A new three-phase, double-nested PCR approach allowed robust melting temperature analysis with enhanced limits of detection (LODs). Finally, newly designed sloppy molecular beacons identified many different mutations using a small number of probes. The assay correctly distinguished wild-type sequences from 32 commonly occurring mutant sequences tested in gyrA, gyrB, katG, and rrs genes and the promoters of inhA and eis genes responsible for resistance to INH, the fluoroquinolone (FQ) drugs, amikacin (AMK), and kanamycin (KAN). The LOD was 300 CFU of Mycobacterium tuberculosis in 1 ml sputum. The rate of detection of heteroresistance by the assay was equivalent to that by Sanger sequencing. In a blind study of 24 clinical sputum samples, resistance mutations were detected in all targets with 100% sensitivity, with the specificity being 93.7 to 100%. Compared to the results of phenotypic susceptibility testing, the sensitivity of the assay was 75% for FQs and 100% each for INH, AMK, and KAN and the specificity was 100% for INH and FQ and 94% for AMK and KAN. Our approach could enable testing for XDR-TB in point-of-care settings, potentially identifying highly drug-resistant TB more quickly and simply than currently available methods.KEYWORDS 10-color assay, three-phase PCR, XDR-TB, point-of-care test

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Rapid Diagnosis of Drug Resistance to Fluoroquinolones, Amikacin, Capreomycin, Kanamycin and Ethambutol Using Genotype MTBDRsl Assay: A Meta-Analysis

Cited by 64 publications

References 51 publications

MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

Molecular Characterization of the Resistance of <i>Mycobacterium tuberculosis</i> to Second Line Drugs in Côte d’Ivoire

Detection of Isoniazid-, Fluoroquinolone-, Amikacin-, and Kanamycin-Resistant Tuberculosis in an Automated, Multiplexed 10-Color Assay Suitable for Point-of-Care Use

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